| 质粒名称: | pCMVInt |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 18935 |
| 存储实验室: | Michele Calos |
|
载体骨架基本信息 |
|
| 载体名称: | pCMVSPORTbetaGal |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4357 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | phiC31 integrase |
| 物种来源: | Streptomyces phage phiC31 |
| 插入大小: | 1873 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SpeI(未破坏) |
| 3' 克隆位点: | BamHI (未破坏) |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCMVInt was a gift from Michele Calos (Addgene plasmid # 18935 ; http://n2t.net/addgene:18935 ; RRID:Addgene_18935) |
| 参考文献部分: | A phage integrase directs efficient site-specific integration in human cells. Groth AC, Olivares EC, Thyagarajan B, Calos MP. Proc Natl Acad Sci U S A. 2000 May 23. 97(11):5995-6000. 10.1073/pnas.090527097 PubMed 10801973 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pCMVInt 6228 bp ds-DNA circular SYN 22-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCMVInt
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6228)
AUTHORS Groth AC, Olivares EC, Thyagarajan B, Calos MP
TITLE A phage integrase directs efficient site-specific integration in
human cells.
JOURNAL Proc Natl Acad Sci U S A. 2000 May 23. 97(11):5995-6000.
PUBMED 10801973
REFERENCE 2 (bases 1 to 6228)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 22, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6228
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 98..115
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 361..379
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 406..436
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 444..460
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 449..471
/label=M13/pUC Reverse
/note="In lacZ gene"
enhancer 490..793
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 794..997
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 948..968
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 994..1018
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 1113..1131
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1113..1130
/label=SP6
/note="SP6 promoter, forward primer"
CDS 1224..3041
/codon_start=1
/product="integrase from phage phi-C31"
/label=phage phi-C31 integrase
/translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
EKRASITWAKPPTDDDEDDAQDGTEDVAA"
primer_bind complement(3104..3123)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3105..3123)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 3510..3644
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3547..3566)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3601..3620
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 3770..4224
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3857..3876)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4066..4087
/label=F1ori-F
/note="F1 origin, forward primer"
CDS 4511..5371
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(4729..4748)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
protein_bind complement(5473..5506)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
rep_origin 5584..6172
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6073..6092
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
181 agcttgcaat tcgcgctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
301 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
361 aataggcgta tcacgaggcc ctttcactca ttaggcaccc caggctttac actttatgct
421 tccggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcat
481 cgtgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
541 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
601 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
661 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
721 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
781 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
841 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
901 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
961 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
1021 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
1081 ccggactcta gcctaggctt ttgcaaaaag ctatttaggt gacactatag aaggtacgcc
1141 tgcaggtacc ggtccggaat tcccgggtcg acgagctcac tagtcgtagg gtcgccgaca
1201 tgacacaagg ggttgtgacc ggggtggaca cgtacgcggg tgcttacgac cgtcagtcgc
1261 gcgagcgcga aaattcgagc gcagcaagcc cagcgacaca gcgtagcgcc aacgaagaca
1321 aggcggccga ccttcagcgc gaagtcgagc gcgacggggg ccggttcagg ttcgtcgggc
1381 atttcagcga agcgccgggc acgtcggcgt tcgggacggc ggagcgcccg gagttcgaac
1441 gcatcctgaa cgaatgccgc gccgggcggc tcaacatgat cattgtctat gacgtgtcgc
1501 gcttctcgcg cctgaaggtc atggacgcga ttccgattgt ctcggaattg ctcgccctgg
1561 gcgtgacgat tgtttccact caggaaggcg tcttccggca gggaaacgtc atggacctga
1621 ttcacctgat tatgcggctc gacgcgtcgc acaaagaatc ttcgctgaag tcggcgaaga
1681 ttctcgacac gaagaacctt cagcgcgaat tgggcgggta cgtcggcggg aaggcgcctt
1741 acggcttcga gcttgtttcg gagacgaagg agatcacgcg caacggccga atggtcaatg
1801 tcgtcatcaa caagcttgcg cactcgacca ctccccttac cggacccttc gagttcgagc
1861 ccgacgtaat ccggtggtgg tggcgtgaga tcaagacgca caaacacctt cccttcaagc
1921 cgggcagtca agccgccatt cacccgggca gcatcacggg gctttgtaag cgcatggacg
1981 ctgacgccgt gccgacccgg ggcgagacga ttgggaagaa gaccgcttca agcgcctggg
2041 acccggcaac cgttatgcga atccttcggg acccgcgtat tgcgggcttc gccgctgagg
2101 tgatctacaa gaagaagccg gacggcacgc cgaccacgaa gattgagggt taccgcattc
2161 agcgcgaccc gatcacgctc cggccggtcg agcttgattg cggaccgatc atcgagcccg
2221 ctgagtggta tgagcttcag gcgtggttgg acggcagggg gcgcggcaag gggctttccc
2281 gggggcaagc cattctgtcc gccatggaca agctgtactg cgagtgtggc gccgtcatga
2341 cttcgaagcg cggggaagaa tcgatcaagg actcttaccg ctgccgtcgc cggaaggtgg
2401 tcgacccgtc cgcacctggg cagcacgaag gcacgtgcaa cgtcagcatg gcggcactcg
2461 acaagttcgt tgcggaacgc atcttcaaca agatcaggca cgccgaaggc gacgaagaga
2521 cgttggcgct tctgtgggaa gccgcccgac gcttcggcaa gctcactgag gcgcctgaga
2581 agagcggcga acgggcgaac cttgttgcgg agcgcgccga cgccctgaac gcccttgaag
2641 agctgtacga agaccgcgcg gcaggcgcgt acgacggacc cgttggcagg aagcacttcc
2701 ggaagcaaca ggcagcgctg acgctccggc agcaaggggc ggaagagcgg cttgccgaac
2761 ttgaagccgc cgaagccccg aagcttcccc ttgaccaatg gttccccgaa gacgccgacg
2821 ctgacccgac cggccctaag tcgtggtggg ggcgcgcgtc agtagacgac aagcgcgtgt
2881 tcgtcgggct cttcgtagac aagatcgttg tcacgaagtc gactacgggc agggggcagg
2941 gaacgcccat cgagaagcgc gcttcgatca cgtgggcgaa gccgccgacc gacgacgacg
3001 aagacgacgc ccaggacggc acggaagacg tagcggcgta gcgagacacc cggatccctc
3061 gaggggccca agcttacgcg tgcatgcgac gtcatagctc tctccctata gtgagtcgta
3121 ttataagcta gcttgggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg
3181 acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat
3241 gtgttaaact agctgcatat gcttgctgct tgagagtttt gcttactgag tatgatttat
3301 gaaaatatta tacacaggag ctagtgattc taattgtttg tgtattttag attcacagtc
3361 ccaaggctca tttcaggccc ctcagtcctc acagtctgtt catgatcata atcagccata
3421 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
3481 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
3541 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
3601 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcgatcct gcattaatga
3661 atcggccaac gcgcggggag aggcggtttg cgtattggct ggcgtaatag cgaagaggcc
3721 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
3781 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
3841 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
3901 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
3961 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
4021 tagacggttt ttcgcccttt gacgttggag tccacgttct taatagtgga ctcttgttcc
4081 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
4141 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
4201 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
4261 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgccag gtcttggact
4321 ggtgagaacg gcttgctcgg cagcttcgat gtgtgctgga gggagaataa aggtctaaga
4381 tgtgcgatag agggaagtcg cattgaatta tgtgctgtgt agggatcgct ggtatcaaat
4441 atgtgtgccc acccctggca tgagacaata accctgataa atgcttcaat aatattgaaa
4501 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
4561 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
4621 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
4681 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
4741 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
4801 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
4861 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
4921 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
4981 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
5041 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
5101 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
5161 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
5221 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
5281 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
5341 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
5401 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
5461 taatctcatg ccataacttc gtataatgta tgctatacga agttatggca tgaccaaaat
5521 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
5581 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
5641 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
5701 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
5761 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
5821 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
5881 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
5941 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
6001 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
6061 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
6121 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
6181 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctca
//
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pCMVInt
2023-07-13
